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  • Eli Boritz
    Junior Member
    • Jul 2010
    • 1
    #1

    oligo-dT Dynabeads wash buffer for mRNA-Seq

    The Dynabeads protocol calls for a wash buffer that includes Lithium chloride, but other published protocols (ex., http://www.ncbi.nlm.nih.gov/pubmed/21807852) replace LiCl with NaCl. I presume this is because Lithium might inhibit RT, but am not sure. Does anyone who does a lot of Illumina TruSeq-based mRNA-seq have a preferred wash buffer for the oligo-dT beads?
    Thanks very much!
    Tags: None
  • mikaelk
    Member
    • Jun 2012
    • 11
    #2
    In the lab we are using a directional mrna-seq protocol based on illumina tru-seq smallrna kits. I have not compared the two types of buffers but I have been doing polyA purifications with LiCl based buffers and I have still been able to construct libraries without any problem. Hope this helps a little.

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    • protist
      Senior Member
      • Jan 2009
      • 101
      #3
      We have used the Li containing buffers and have had no issues with libraries - even from small amounts of starting material.

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